在哺乳动物细胞中使用构成性活性转录因子VP16-CREB缓解转录瓶
Chansik Yoon1, Kyoung Eun Baek1, Dongil Kim1
1Department of Biological Sciences, KAIST, Daejeon, 34141, Republic of Korea.
Metabolic engineering
|September 14, 2023
概括
这项研究设计了转录因子 (TFs),以促进哺乳动物细胞中的重组蛋白质生产. 构成性活跃的CREB显著增加了治疗性蛋白质产量,克服了转录的极限.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
背景情况:
- 哺乳动物细胞中高水平的重组蛋白表达对治疗至关重要.
- 转录效率低下是限制蛋白质产量的一个主要瓶.
- 由于转录因子 (TF) 的可用性,现有的合成促进剂显示出有限的成功.
研究的目的:
- 开发一种TF工程策略,以克服重组蛋白生产中的转录瓶.
- 识别和利用特定的TF来提高哺乳动物细胞的转录效率.
- 为了提高复合中国仓鼠卵巢 (rCHO) 细胞系中治疗性蛋白质的产量.
主要方法:
- 转录组分析确定了cAMP响应元素结合蛋白 (CREB) 作为高生产rCHO细胞中的候选TF.
- 在rCHO细胞系中评估了活性CREB的转录限制.
- 表达的构成性活性VP16-CREB直接激活转基因促进体在非细胞毒性水平.
主要成果:
- 在CMV促体驱动的rCHO细胞中,VP16-CREB的表达增加了治疗性蛋白质的产生 (单克隆抗体,乙素) 高达3.9倍.
- VP16-CREB成功激活了含有cAMP反应元素的合成促进剂.
- 证明构成性活跃的TFs可以克服重组哺乳动物系统中的转录限制.
结论:
- TF工程提供了一种可行的策略,以增强哺乳动物细胞中的重组蛋白质表达.
- 构成性活性的CREB有效地提高了治疗性蛋白质的产量.
- 这种方法提供了一种缓解转录瓶的方法,以改善生物制造.
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